GSM8046684: RGC, experiment4, GEX, scRNA-seq; Mus musculus; RNA-Seq
Fresh mouse retinas were dissected out from Vglut2-Tdtomato-Ai9 mice in oxygenated Ames media. Retinas are digested in papain and dissociated into single cells by manual trituration in low-ovomucoid solution. RGCs were enriched by negative-rod immunopanning using an anti-CD73 antibody. RGCs were labeled with a Thy1-APC antibody. RGCs were incubated with cholesteral-modified oligos using the 10x Genomics 3' CellPlex Kit (PN-1000261). RGCs from up to 6 retinas are pooled and resuspended in oxygenated Ames media, then enriched using FACS to isolate APC and Tdtomato positive cells. Unless specified, all steps were performed in oxygenated Ames, in the presence of Actinomycin D. All steps were also performed on individual retinas up until FACS. Library construction was performed according to the manufacturer's instructions for the Chromium Next GEM 3' Kit v3.1